Denaturing Gradient Gel Electrophoresis

Denaturing gradient gel electrophoresis (DGGE) separates DNA molecules differing in sequence by only a single base. As its name indicates, this technique combines gel electrophoresis with DNA denaturation. It is applicable to double-stranded DNA fragments of a few hundred bases, which are often generated by PCR (see Ch. 23). During DGGE, double-stranded DNA is subjected to a gradient of increasing denaturation to separate the strands. The dsDNA melts in stages in which discrete zones known as "melting domains", become unpaired along the piece of DNA. Partially melted DNA migrates more slowly through an agarose gel during electrophoresis because the small streamlined DNA fragments open into larger structures that get denaturing gradient gel electrophoresis (DGGE) Combination of gel electrophoresis with DNA denaturation that allows separation of DNA molecules differing in sequence by only a single base

Denaturing Gradient Gel Electrophoresis 573

kilobases 1,500

1,000 800 600

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